N cadherin 610920

Welgene Inc. (Seoul, Korea) supplied the Roswell Park Memorial Institute (RPMI)-1640 medium, fetal bovine serum (FBS), phosphate-buffered saline (PBS), and penicillin and streptomycin (PS). Lipofectamine™ 2000 or JetPEI was supplied by Invitrogen (Carlsbad, CA, USA) or Polyplus-transfection (Illkirch-Graffenstaden, France), respectively. Sphingosylphosphorylcholine (SPC) was procured from Matreya (Pleasant Gap, PA, USA). Calbiochem (La Jolla, CA, USA) supplied the MAP kinase inhibitors such as PD98059, SP600125, and SB203580. Mouse monoclonal antibodies specific for E-cadherin (610181, 1:1000), N-cadherin (610920, 1:1000), and PP2A (610555, 1:2000) were from BD Biosciences (San Jose, CA, USA). The anti-YDJC (ab107873, 1:500) antibody was supplied by Abcam (Cambridge, MA, USA). Cell Signalling Technology (Beverly, MA, USA) provided the phosphospecific antibody to detect ERK1/2 (#9102, 1:1000) and anti-ERK (#9106, 1:1000) antibodies. Anti-ubiquitin, anti-CDC16 (sc-365636, 1:1000), and anti-β-actin (sc-47778, 1:5000) antibodies were provided by Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA). Molecular Probes, Inc. (Eugene, OR, USA) supplied Alexa Fluor 594 goat anti-mouse and Alexa Fluor 488 goat anti-rabbit antibodies. All of these reagents were freshly made at the moment of each experiment.

API (A3145) and dimethyl sulfoxide (DMSO) were purchased from Sigma-Aldrich (St. Louis, MO). Fetal bovine serum (FBS), antibiotics, molecular weight standards, trypsin-EDTA, and all medium additives were obtained from Life Technologies (Gaithersburg, MD). The CellTiter 96 AQueous One Solution Proliferation Assay System was purchased from Promega (Madison, WI). An enhanced chemiluminescence (ECL) kit was purchased from Amersham (Arlington Heights, IL). Antibodies specific for fibronectin (ab2413) and SPOCK1 (ab229935) were obtained from Abcam (Cambridge, MA). Antibodies specific for cleaved-PARP (#9541), phospho-Ak (ser473, #9271), Akt (#4691), Snail (#3895), Slug (#9585), Twist (#46702), and E-cadherin (#3195) were obtained from Cell Signaling Technology (Danvers, MA). Antibodies specific for vimentin (550513) and N-cadherin (610920) were purchased from BD Biosciences (San Jose, CA). Antibodies specific for cyclin D1 (sc-8396), cyclin E (sc-377,100), β-actin (sc-47,778), and goat anti-rabbit (sc-2004) and anti-mouse (sc-2005) immunoglobulin G (IgG) antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA). Polyvinylidene fluoride (PVDF) membranes for Western blotting were purchased from Bio-Rad (Hercules, CA). Unless otherwise specified, other chemicals used in this study were purchased from Sigma Chemical (St. Louis, MO).